钱小兰, 帅怡, 王彦琴, 肖萍, 仲伟鉴. 微囊藻毒素-LR对原代大鼠肝细胞p-JNK1/2、p-P44/42、p-P38蛋白水平的影响[J]. 环境与职业医学, 2013, 30(5): 356-359,364.
引用本文: 钱小兰, 帅怡, 王彦琴, 肖萍, 仲伟鉴. 微囊藻毒素-LR对原代大鼠肝细胞p-JNK1/2、p-P44/42、p-P38蛋白水平的影响[J]. 环境与职业医学, 2013, 30(5): 356-359,364.
QIAN Xiao-lan , SHUAI Yi , WANG Yan-Qin , XIAO Ping , ZHONG Wei-jian . Microcystin-LR Induces Phosphorylation of p-JNK1/2, P44/42, P38 in Primary Cultured Rat Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2013, 30(5): 356-359,364.
Citation: QIAN Xiao-lan , SHUAI Yi , WANG Yan-Qin , XIAO Ping , ZHONG Wei-jian . Microcystin-LR Induces Phosphorylation of p-JNK1/2, P44/42, P38 in Primary Cultured Rat Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2013, 30(5): 356-359,364.

微囊藻毒素-LR对原代大鼠肝细胞p-JNK1/2、p-P44/42、p-P38蛋白水平的影响

Microcystin-LR Induces Phosphorylation of p-JNK1/2, P44/42, P38 in Primary Cultured Rat Hepatocytes

  • 摘要: 目的 研究低剂量微囊藻毒素-LR(microcystin-LR, MC-LR)染毒原代大鼠肝细胞对细胞内磷酸化的c-jun 氨基末端激酶(p-JNK1/2)、磷酸化的细胞外信号激酶(p-P44/42)、磷酸化的P38 丝裂原活化蛋白激酶(p-P38)蛋白水平的影响。

    方法 采用改良胶原酶灌流法建立的原代大鼠肝细胞培养模型, 以MC-LR 终浓度1& #215;10-9、1& #215;10-10、1& #215;10-11、1& #215;10-12 mol/L 4 个剂量染毒细胞, 分别培养至3、6、12、24 h 4 个时间点后裂解细胞获取总蛋白, 用Western 法检测分析丝裂原蛋白激酶家族(MAPKs)磷酸化蛋白p-JNK1/2、p-P44/42、p-P38 水平。

    结果 染毒原代大鼠肝细胞3、6 h 后p-JNK1/2、p-P44/42、p-P38 蛋白表达水平随染毒剂量降低而上升, 在1& #215;10-10 或1& #215;10-11 mol/L 剂量组蛋白水平达到峰值后随染毒剂量降低而蛋白水平增幅减小;染毒至12 h 除p-P38 各剂量组蛋白水平升高外, 其余目标蛋白各剂量组与对照组相比差异无统计学意义;染毒24 h 各目标蛋白各剂量组与对照组相比差异无统计学意义。

    结论 低剂量微囊藻毒素可促进原代大鼠肝细胞p-JNK1/2、p-P44/42、p-P38 水平升高且具有时间效应关系。

     

    Abstract: Objective To investigate the effect of low-dose microcystin-LR(MC-LR)on phosphorylation of JNK1/2, P44/42, and P38 in primary cultured rat hepatocytes.

    Methods Primary rat hepatocytes were isolated by collagenase perfusion method. The obtained cells were treated with 1& #215;10-9, 1& #215;10-10, 1& #215;10-11, 1& #215;10-12 mol/L MC-LR for 3, 6, 12, and 24 h, respectively, then harvested and lysed by radio immunoprecipitation assay(RIPA). The expressions of p-JNK1/2, p-P44/42, and p-P38 were detected by Western blot.

    Results With the decrease in dose of MC-LR, the expressions of p-JNK1/2, p-P44/42, and p-P38 increased after 3 h and 6 h of incubation across all designed concentrations of MC-LR, and a peak was observed at 1& #215;10-10 or 1& #215;10-11 mol/L, then the effect intensity decreased with the dose reduction. After the cells exposed for 12 h, only p-P38 showed a slight rise, and no significant differences in phosphorylation expression were found between the various dose groups and the control group. No outstanding effect was observed after 24 h exposure to MC-LR.

    Conclusion Low-dose of MC-LR could enhance the phosporylation of JNK1/2, P44/42, and P38, and the effect exhibits time-response relationship.

     

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